Herpes Simplex Virus Causes Amplification of Recombinant Plasmids Containing Simian Virus 40 Sequences

Matz, Bertfried

doi:10.1099/0022-1317-70-6-1347

Cited by 17 publications

(16 citation statements)

References 48 publications

(40 reference statements)

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“…It has been shown that SV40 origin-containing plasmids replicate in hamster cells upon infection with HSV (Matz, 1989). This heterologous SV40 DNA synthesis is mediated by the HSV replication genes (UL5, UL8, UL29, UL30, UL42 and UL52; Heilbronn & zur Hausen, 1989) but does not require the UL9 gene (Heilbronn & zur Hausen, 1989;Matz, 1989). It seems that T antigen substitutes for UL9 protein on SV40 DNA and makes the SV40 DNA accessible to the other HSV-specific DNA replication proteins.…”

Section: Orf (Mcgeochmentioning

confidence: 99%

“…Transfection and analysis of plasmid replication were carried out as previously described (Matz, 1989). It has been shown that SV40 origin-containing plasmids replicate in hamster cells upon infection with HSV (Matz, 1989). This heterologous SV40 DNA synthesis is mediated by the HSV replication genes (UL5, UL8, UL29, UL30, UL42 and UL52; Heilbronn & zur Hausen, 1989) but does not require the UL9 gene (Heilbronn & zur Hausen, 1989;Matz, 1989).…”

Section: Orf (Mcgeochmentioning

confidence: 99%

Section: Orf (Mcgeochmentioning

confidence: 99%

“…In these experiments baby hamster kidney (BHK) cells were transiently cotransfected with the HSV oti scontaining plasmid pFR106 (Matz, 1989) and SV40 origin-containing plasmid pJS36 (nucleotides 2533 to 274 of SV40 strain VA4554 in pUC18). Transfection and analysis of plasmid replication were carried out as previously described (Matz, 1989).…”

Section: Orf (Mcgeochmentioning

confidence: 99%

“…From parallel cell cultures, DNA was also harvested at times corresponding to the temperature shifts (right panels). The DNA was cleaved with Kpnl and DpnI and plasmid DNA was detected by Southern blot analysis using a a2P-labelled vector probe as described previously (Matz, 1989). Lane M contains a DNA size marker.…”

Section: Orf (Mcgeochmentioning

confidence: 99%

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Thermosensitive UL9 gene function is required for early stages of herpes simplex virus type 1 DNA synthesis

Blümel

Matz²

1995

Journal of General Virology

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DNA replication of herpes simplex virus type 1 (HSV-1) is dependent on a virus-encoded sequence-specific origin-binding protein, the product of the UL9 reading frame. We have identified the mutations in the UL9 gene of three temperature-sensitive (ts) mutants of HSV-1 which are responsible for the ts phenotype (A90T in mutant tsS and V220M in tsR and tsX). The mutations are located in two different conserved helicase sequence motifs of UL9. Two further alterations (I204T and E280D) compared to the published sequence were found in the mutant, revertant and parental wild-type strain 17syn + sequences and therefore seemed to be irrelevant for the ts phenotype. The ts function of the UL9 protein was required at early times during DNA synthesis whereas upward temperature shifts at later times did not considerably inhibit DNA synthesis.Herpes simplex virus type 1 (HSV-1) is able to infect productively many cell types. The virus genome encodes several proteins involved in nucleic acid metabolism. Seven virus genes are necessary and sufficient for amplification of plasmids bearing HSV-specific origins of replication in transiently transfected cells. These genes encode a two subunit DNA polymerase (UL30 and UL42), a ssDNA-binding protein ICP8 (UL29), a three protein complex with helicase/primase activities (UL5, UL8 and UL52) and an HSV origin-specific DNAbinding protein (UL9) (for review see Challberg, 1991).Since the UL9 protein is able to bind specifically to the virus origin of replication, it has been proposed as an initiator protein for HSV DNA replication. The interaction of UL9 protein with its recognition sequence in the virus origins has been investigated in detail (Challberg, 1991). In addition to its DNA-binding property, UL9 also functions as a DNA helicase (Boehmer et al., 1993;Bruckner et al., 1991;Fierer & Challberg, 1992) and conserved helicase motifs within UL9 are essential for DNA replication (Martinez et al., 1992 is believed that UL9 specifically unwinds the virus origin of replication, although this activity has not yet been demonstrated (Fierer & Challberg, 1992;Boehmer et al., 1993). Though a considerable amount of evidence has accumulated that origin-binding of UL9 is crucial for virus DNA replication, a direct role for UL9 in initiation is based on analogy to other systems of DNA replication. To investigate the function of UL9 in vivo we have analysed temperature-sensitive (ts) HSV mutants tsS, tsR and tsX (Matz et al., 1983). This type of mutant has been used in many other systems of DNA replication to characterize the in vivo function of the proteins involved. As it is possible to inactivate the ts function in vivo by an increase in incubation temperature, we have investigated the temporal requirement for UL9 function during replication of HSV origin of replication (oris)-Containing test plasmids. Our results show that the ts function of UL9 is required in the initial phase of DNA replication and are consistent with a model in which UL9 acts primarily during the early stages of DNA synthesis....

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Section: Orf (Mcgeochmentioning

confidence: 99%

Section: Orf (Mcgeochmentioning

confidence: 99%

Section: Orf (Mcgeochmentioning

confidence: 99%

Section: Orf (Mcgeochmentioning

confidence: 99%

Section: Orf (Mcgeochmentioning

confidence: 99%

See 3 more Smart Citations

Thermosensitive UL9 gene function is required for early stages of herpes simplex virus type 1 DNA synthesis

Blümel

Matz²

1995

Journal of General Virology

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Transformation of rodent fibroblasts by herpes simplex virus: Presence of morphological transforming region 1 (MTR 1) is not required for the maintenance of the transformed state

Bauer

Kahl

Sawhney

et al. 1992

Intl Journal of Cancer

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Studies on the mechanisms of transformation of mammalian cells by herpes simplex virus (HSV) in vitro have been prevented so far by the extremely low transformation frequencies obtained in monolayer culture. Here we present a transformation system that relies on the direct seeding in soft agar of infected single cells, thus avoiding negative interactions between normal and transformed cells. We took advantage of HSV-I temperature-sensitive mutants at the UL9 locus, which codes for a DNA-binding protein necessary for viral DNA replication. At the non-permissive temperature, viral DNA synthesis and late gene expression are prevented. Viral gene expression is restricted to immediate early and early genes. Induction of transformation was highly efficient in our one-step transformation system. It depended on intact viral particles and viral DNA. Immediate early and/or early viral gene expression was sufficient to induce transformation. Colonies were stably transformed and did not show any rescue of viable virus after temperature downshift and co-cultivation with susceptible cells. Transformed cells maintained the transformed state in the absence of viral DNA. Our data therefore support the "hit-and-run" hypothesis for the transforming effect of HSV.

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Structure of Simian Virus 40 DNA Replicated by Herpes Simplex Virus Type 1

2000

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Replicating herpes simplex virus type 1 (HSV-1) DNA is known to form large branched structures. The aim of this study was to define whether HSV-1-specific DNA elements in cis play a critical role in formation of this structure. We did this by investigating the structure of heterologous simian virus 40 (SV40) DNA, which is replicated in HSV-infected cells by SV40 large T-antigen and defined HSV-encoded replication factors (e.g., DNA polymerase, single-stranded DNA-binding protein, and helicase-primase). During this process, extrachromosomal concatemeric DNA replication products are formed, indicating a herpesvirus-specific replication mode. In this study, we found that the replicating SV40 DNA consisted of a complex branched structure indistinguishable from that of replicating HSV DNA. Thus, no HSV-specific DNA element is necessary in cis for the formation of the large branched structure during HSV DNA replication. The trans-acting HSV DNA replication proteins seem to be sufficient to generate these complex structures. Moreover, replicating SV40 DNA showed a high frequency of homologous recombination events, which is typical for HSV DNA replication. However, in contrast to HSV origin-bearing amplicon plasmids, SV40 plasmids bearing the HSV cleavage-packaging signal were not efficiently processed to linear 150-kb DNA packaged into HSV capsids. This indicates that initiation of DNA synthesis on HSV-ori determines some, yet undefined, property of replicating HSV DNA, which is crucial for regular processing of the replication intermediates to daughter genomes.

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Herpes Simplex Virus Causes Amplification of Recombinant Plasmids Containing Simian Virus 40 Sequences

Cited by 17 publications

References 48 publications

Thermosensitive UL9 gene function is required for early stages of herpes simplex virus type 1 DNA synthesis

Thermosensitive UL9 gene function is required for early stages of herpes simplex virus type 1 DNA synthesis

Transformation of rodent fibroblasts by herpes simplex virus: Presence of morphological transforming region 1 (MTR 1) is not required for the maintenance of the transformed state

Structure of Simian Virus 40 DNA Replicated by Herpes Simplex Virus Type 1

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